All Title Author
Keywords Abstract

PLOS ONE  2012 

EphA3 Expressed in the Chicken Tectum Stimulates Nasal Retinal Ganglion Cell Axon Growth and Is Required for Retinotectal Topographic Map Formation

DOI: 10.1371/journal.pone.0038566

Full-Text   Cite this paper   Add to My Lib


Background Retinotopic projection onto the tectum/colliculus constitutes the most studied model of topographic mapping and Eph receptors and their ligands, the ephrins, are the best characterized molecular system involved in this process. Ephrin-As, expressed in an increasing rostro-caudal gradient in the tectum/colliculus, repel temporal retinal ganglion cell (RGC) axons from the caudal tectum and inhibit their branching posterior to their termination zones. However, there are conflicting data regarding the nature of the second force that guides nasal axons to invade and branch only in the caudal tectum/colliculus. The predominant model postulates that this second force is produced by a decreasing rostro-caudal gradient of EphA7 which repels nasal optic fibers and prevents their branching in the rostral tectum/colliculus. However, as optic fibers invade the tectum/colliculus growing throughout this gradient, this model cannot explain how the axons grow throughout this repellent molecule. Methodology/Principal Findings By using chicken retinal cultures we showed that EphA3 ectodomain stimulates nasal RGC axon growth in a concentration dependent way. Moreover, we showed that nasal axons choose growing on EphA3-expressing cells and that EphA3 diminishes the density of interstitial filopodia in nasal RGC axons. Accordingly, in vivo EphA3 ectodomain misexpression directs nasal optic fibers toward the caudal tectum preventing their branching in the rostral tectum. Conclusions We demonstrated in vitro and in vivo that EphA3 ectodomain (which is expressed in a decreasing rostro-caudal gradient in the tectum) is necessary for topographic mapping by stimulating the nasal axon growth toward the caudal tectum and inhibiting their branching in the rostral tectum. Furthermore, the ability of EphA3 of stimulating axon growth allows understanding how optic fibers invade the tectum growing throughout this molecular gradient. Therefore, opposing tectal gradients of repellent ephrin-As and of axon growth stimulating EphA3 complement each other to map optic fibers along the rostro-caudal tectal axis.


[1]  Scicolone G, Ortalli AL, Carri NG (2009) Key roles of Ephs and ephrins in retinotectal topographic map formation. Brain Res Bull 79: 227–247.
[2]  Feldheim DA, O'Leary DD (2010) Visual map development: bidirectional signaling, bifunctional guidance molecules, and competition. Cold Spring Harb Perspect Biol 2: a001768.
[3]  Rapacioli M, Rodriguez Celin A, Duarte S, Ortalli AL, Di Napoli J, et al. (2011) The chick optic tectum developmental stages. A dynamic table based on temporal- and spatial-dependent histogenetic changes: A structural, morphometric and immunocytochemical analysis. J Morphol 272: 675–697.
[4]  Scicolone G, Ortalli AL, Alvarez G, Lopez-Costa JJ, Rapacioli M, et al. (2006) Developmental pattern of NADPH-diaphorase positive neurons in chick optic tectum is sensitive to changes in visual stimulation. J Comp Neurol 494: 1007–1030.
[5]  Scicolone G, Pereyra-Alfonso S, Brusco A, Pecci Saavedra J, Flores V (1995) Development of the laminated pattern of the chick tectum opticum. Int J Dev Neurosci 13: 845–858.
[6]  Yates PA, Roskies AL, McLaughlin T, O'Leary DD (2001) Topographic-specific axon branching controlled by ephrin-As is the critical event in retinotectal map development. J Neurosci 21: 8548–8563.
[7]  Sperry RW (1963) Chemoaffinity in the Orderly Growth of Nerve Fiber Patterns and Connections. Proc Natl Acad Sci U S A 50: 703–710.
[8]  Gebhardt C, Bastmeyer M, Weth F (2012) Balancing of ephrin/Eph forward and reverse signaling as the driving force of adaptive topographic mapping. Development 139: 335–345.
[9]  Bevins N, Lemke G, Reber M (2011) Genetic Dissection of EphA Receptor Signaling Dynamics during Retinotopic Mapping. J Neurosci 31: 10302–10310.
[10]  Reber M, Burrola P, Lemke G (2004) A relative signalling model for the formation of a topographic neural map. Nature 431: 847–853.
[11]  Pfeiffenberger C, Yamada J, Feldheim DA (2006) Ephrin-As and patterned retinal activity act together in the development of topographic maps in the primary visual system. J Neurosci 26: 12873–12884.
[12]  Simpson HD, Mortimer D, Goodhill GJ (2009) Theoretical models of neural circuit development. Curr Top Dev Biol 87: 1–51.
[13]  Tsigankov D, Koulakov AA (2010) Sperry versus Hebb: topographic mapping in Isl2/EphA3 mutant mice. BMC Neurosci 11: 155.
[14]  Flanagan JG (2006) Neural map specification by gradients. Curr Opin Neurobiol 16: 59–66.
[15]  McLaughlin T, O'Leary DD (2005) Molecular gradients and development of retinotopic maps. Annu Rev Neurosci 28: 327–355.
[16]  Pasquale EB (2005) Eph receptor signalling casts a wide net on cell behaviour. Nat Rev Mol Cell Biol 6: 462–475.
[17]  Vearing CJ, Lackmann M (2005) “Eph receptor signalling; dimerisation just isn't enough”. Growth Factors 23: 67–76.
[18]  Connor RJ, Menzel P, Pasquale EB (1998) Expression and tyrosine phosphorylation of Eph receptors suggest multiple mechanisms in patterning of the visual system. Dev Biol 193: 21–35.
[19]  Menzel P, Valencia F, Godement P, Dodelet VC, Pasquale EB (2001) Ephrin-A6, a new ligand for EphA receptors in the developing visual system. Dev Biol 230: 74–88.
[20]  Brown A, Yates PA, Burrola P, Ortuno D, Vaidya A, et al. (2000) Topographic mapping from the retina to the midbrain is controlled by relative but not absolute levels of EphA receptor signaling. Cell 102: 77–88.
[21]  Carreres MI, Escalante A, Murillo B, Chauvin G, Gaspar P, et al. (2011) Transcription factor Foxd1 is required for the specification of the temporal retina in mammals. J Neurosci 31: 5673–5681.
[22]  Park S, Frisen J, Barbacid M (1997) Aberrant axonal projections in mice lacking EphA8 (Eek) tyrosine protein kinase receptors. Embo J 16: 3106–3114.
[23]  Marin O, Blanco MJ, Nieto MA (2001) Differential expression of Eph receptors and ephrins correlates with the formation of topographic projections in primary and secondary visual circuits of the embryonic chick forebrain. Dev Biol 234: 289–303.
[24]  Rashid T, Upton AL, Blentic A, Ciossek T, Knoll B, et al. (2005) Opposing gradients of ephrin-As and EphA7 in the superior colliculus are essential for topographic mapping in the mammalian visual system. Neuron 47: 57–69.
[25]  Hornberger MR, Dutting D, Ciossek T, Yamada T, Handwerker C, et al. (1999) Modulation of EphA receptor function by coexpressed ephrinA ligands on retinal ganglion cell axons. Neuron 22: 731–742.
[26]  Monschau B, Kremoser C, Ohta K, Tanaka H, Kaneko T, et al. (1997) Shared and distinct functions of RAGS and ELF-1 in guiding retinal axons. Embo J 16: 1258–1267.
[27]  Cheng HJ, Nakamoto M, Bergemann AD, Flanagan JG (1995) Complementary gradients in expression and binding of ELF-1 and Mek4 in development of the topographic retinotectal projection map. Cell 82: 371–381.
[28]  Drescher U, Kremoser C, Handwerker C, Loschinger J, Noda M, et al. (1995) In vitro guidance of retinal ganglion cell axons by RAGS, a 25 kDa tectal protein related to ligands for Eph receptor tyrosine kinases. Cell 82: 359–370.
[29]  Feldheim DA, Kim YI, Bergemann AD, Frisen J, Barbacid M, et al. (2000) Genetic analysis of ephrin-A2 and ephrin-A5 shows their requirement in multiple aspects of retinocollicular mapping. Neuron 25: 563–574.
[30]  Nakamoto M, Cheng HJ, Friedman GC, McLaughlin T, Hansen MJ, et al. (1996) Topographically specific effects of ELF-1 on retinal axon guidance in vitro and retinal axon mapping in vivo. Cell 86: 755–766.
[31]  Sakurai T, Wong E, Drescher U, Tanaka H, Jay DG (2002) Ephrin-A5 restricts topographically specific arborization in the chick retinotectal projection in vivo. Proc Natl Acad Sci U S A 99: 10795–10800.
[32]  Feldheim DA, Nakamoto M, Osterfield M, Gale NW, DeChiara TM, et al. (2004) Loss-of-function analysis of EphA receptors in retinotectal mapping. J Neurosci 24: 2542–2550.
[33]  Carvalho RF, Beutler M, Marler KJ, Knoll B, Becker-Barroso E, et al. (2006) Silencing of EphA3 through a cis interaction with ephrinA5. Nat Neurosci 9: 322–330.
[34]  Dutting D, Handwerker C, Drescher U (1999) Topographic targeting and pathfinding errors of retinal axons following overexpression of ephrinA ligands on retinal ganglion cell axons. Dev Biol 216: 297–311.
[35]  Gosse NJ, Nevin LM, Baier H (2008) Retinotopic order in the absence of axon competition. Nature 452: 892–895.
[36]  Lim YS, McLaughlin T, Sung TC, Santiago A, Lee KF, et al. (2008) p75 (NTR) mediates ephrin-A reverse signaling required for axon repulsion and mapping. Neuron 59: 746–758.
[37]  Marler KJ, Becker-Barroso E, Martinez A, Llovera M, Wentzel C, et al. (2008) A TrkB/EphrinA interaction controls retinal axon branching and synaptogenesis. J Neurosci 28: 12700–12712.
[38]  Marler KJ, Poopalasundaram S, Broom ER, Wentzel C, Drescher U (2010) Pro-neurotrophins secreted from retinal ganglion cell axons are necessary for ephrinA-p75NTR-mediated axon guidance. Neural Dev 5: 30.
[39]  Poopalasundaram S, Marler KJ, Drescher U (2011) EphrinA6 on chick retinal axons is a key component for p75(NTR)-dependent axon repulsion and TrkB-dependent axon branching. Mol Cell Neurosci 47: 131–136.
[40]  McLaughlin T, Hindges R, O'Leary DD (2003) Regulation of axial patterning of the retina and its topographic mapping in the brain. Curr Opin Neurobiol 13: 57–69.
[41]  Herrera E, Marcus R, Li S, Williams SE, Erskine L, et al. (2004) Foxd1 is required for proper formation of the optic chiasm. Development 131: 5727–5739.
[42]  Takahashi H, Sakuta H, Shintani T, Noda M (2009) Functional mode of FoxD1/CBF2 for the establishment of temporal retinal specificity in the developing chick retina. Dev Biol 331: 300–310.
[43]  Takahashi H, Shintani T, Sakuta H, Noda M (2003) CBF1 controls the retinotectal topographical map along the anteroposterior axis through multiple mechanisms. Development 130: 5203–5215.
[44]  Muhleisen TW, Agoston Z, Schulte D (2006) Retroviral misexpression of cVax disturbs retinal ganglion cell axon fasciculation and intraretinal pathfinding in vivo and guidance of nasal ganglion cell axons in vivo. Dev Biol 297: 59–73.
[45]  Cohen J, Nurcombe V, Jeffrey P, Edgar D (1989) Developmental loss of functional laminin receptors on retinal ganglion cells is regulated by their target tissue, the optic tectum. Development 107: 381–387.
[46]  Halfter W, Claviez M, Schwarz U (1981) Preferential adhesion of tectal membranes to anterior embryonic chick retina neurites. Nature 292: 67–70.
[47]  Wang HU, Anderson DJ (1997) Eph family transmembrane ligands can mediate repulsive guidance of trunk neural crest migration and motor axon outgrowth. Neuron 18: 383–396.
[48]  Davenport RW, Thies E, Cohen ML (1999) Neuronal growth cone collapse triggers lateral extensions along trailing axons. Nat Neurosci 2: 254–259.
[49]  Dent EW, Kalil K (2001) Axon branching requires interactions between dynamic microtubules and actin filaments. J Neurosci 21: 9757–9769.
[50]  Bonhoeffer F, Huf J (1985) Position-dependent properties of retinal axons and their growth cones. Nature 315: 409–410.
[51]  Kao TJ, Kania A (2011) Ephrin-Mediated cis-Attenuation of Eph Receptor Signaling Is Essential for Spinal Motor Axon Guidance. Neuron 71: 76–91.
[52]  Yoo S, Kim Y, Noh H, Lee H, Park E, et al. (2011) Endocytosis of EphA receptors is essential for the proper development of the retinocollicular topographic map. Embo J 30: 1593–1607.
[53]  Brunet I, Weinl C, Piper M, Trembleau A, Volovitch M, et al. (2005) The transcription factor Engrailed-2 guides retinal axons. Nature 438: 94–98.
[54]  Wizenmann A, Brunet I, Lam JS, Sonnier L, Beurdeley M, et al. (2009) Extracellular Engrailed participates in the topographic guidance of retinal axons in vivo. Neuron 64: 355–366.
[55]  Matsunaga E, Nakamura H, Chedotal A (2006) Repulsive guidance molecule plays multiple roles in neuronal differentiation and axon guidance. J Neurosci 26: 6082–6088.
[56]  Hamburger V, Hamilton H (1951) A series of normal stages in the development of the chick embryo. J Morphol. pp. 49–92.
[57]  Hughes SH, Greenhouse JJ, Petropoulos CJ, Sutrave P (1987) Adaptor plasmids simplify the insertion of foreign DNA into helper-independent retroviral vectors. J Virol 61: 3004–3012.
[58]  Fekete DM, Cepko CL (1993) Replication-competent retroviral vectors encoding alkaline phosphatase reveal spatial restriction of viral gene expression/transduction in the chick embryo. Mol Cell Biol 13: 2604–2613.
[59]  Jaskolski F, Mulle C, Manzoni OJ (2005) An automated method to quantify and visualize colocalized fluorescent signals. J Neurosci Methods 146: 42–49.
[60]  Zinchuk V, Grossenbacher-Zinchuk O (2009) Recent advances in quantitative colocalization analysis: focus on neuroscience. Prog Histochem Cytochem 44: 125–172.
[61]  Cowan WM, Adamson L, Powell TP (1961) An experimental study of the avian visual system. J Anat 95: 545–563.


comments powered by Disqus